Timmermans M C, Das O P, Messing J
Waksman Institute, Rutgers, State University of New Jersey, Piscataway 08855-0759.
Nucleic Acids Res. 1992 Aug 11;20(15):4047-54. doi: 10.1093/nar/20.15.4047.
The replication of shuttle vectors derived from Wheat Dwarf Virus, a monopartite geminivirus, was studied in cultured maize endosperm cells, and in the Black Mexican Sweet (BMS) maize cell line. Using in vivo labeling and DNA methylation analysis, we showed that replication was initiated within 24 hrs after transfection, and did not require cell division in both cell lines. Copy numbers of 30,000 ds DNA molecules per cell were observed in endosperm cells after three days. The replication protein was shown to act in trans, since the wild type gene of the shuttle vector enabled replication-deficient vectors carrying mutated genes to replicate. These properties suggest that WDV may have similar applications in plants as SV40 in mammalian cells.
从小麦矮缩病毒（一种单分体双生病毒）衍生而来的穿梭载体在培养的玉米胚乳细胞和黑色墨西哥甜玉米（BMS）细胞系中的复制情况得到了研究。通过体内标记和DNA甲基化分析，我们发现转染后24小时内即可启动复制，并且在这两种细胞系中复制均不需要细胞分裂。三天后，在胚乳细胞中观察到每个细胞有30,000个双链DNA分子的拷贝数。由于穿梭载体的野生型基因能使携带突变基因的复制缺陷型载体进行复制，因此表明复制蛋白具有反式作用。这些特性表明，WDV在植物中的应用可能与SV40在哺乳动物细胞中的应用类似。
